A hyperspectral microarray scanner and a hyperspectral confocal microscope have been developed over the past several years. See M. B. Sinclair et al., Applied Optics 43, 2079 (2004); and M. B. Sinclair et al., Applied Optics 45, 6283 (2006). In addition, efficient multivariate data analysis algorithms and software have been developed. See J. A. T. Ohlhausen et al., Applied Surface Science 231/232, 230 (2004); H. D. T. Jones et al., J. Chemometrics 22, 482 (2008); and D. M. Haaland et al., Appl. Spectrosc. 63, 271 (2009). Initial application of these hyperspectral technologies to biological investigations has revealed several distinct advantages of the hyperspectral approach, including: improved accuracy for the measurement of low level emissions; the ability to simultaneously monitor many emitting species; the ability to identify and remove unwanted contributions from contaminants; and the ability to identify and utilize (or remove) cell autofluorescence. See J. A. Timlin et al., Proc. SPIE 6088, 608805-1 (2006); M. J. Martinez et al., Nucleic Acids Research 31, 1 (2003); J. A. Timlin et al., Bmc Genomics 6, 72 (2005); J. A. Timlin et al., IEEE International Symposium on Biomedical Imaging, 1529 (2004); and J. A. Timlin et al., Proc. SPIE 6859, 68590A-1 (2008). More recently, the advantages of these hyperspectral imaging technologies have been clearly demonstrated in a series of detailed biological investigations. See R. W. Davis et al., Microscopy and Microanalysis 16, 478 (2010); R. W. Davis et al., Journal of Peptide Science 15, 511 (2009); W. F. J. Vermaas et al., Proceedings of the National Academy of Sciences of the United States of America 105, 4050 (2008); V. L. Sutherland et al., Journal of Neuroscience Methods 160, 144 (2007); and M. C. Pedroso et al., Microscopy Today 18, 14 (2010).
During this same time period, the bioscience community has placed increasing emphasis on the development of high throughput instrumentation. The general advantages of increased throughput include the ability to characterize a larger number of compounds, as well as an increase in the reliability and statistical significance of the measurements. Thus, in genomics and proteomics, increased throughput allows for more accurate investigation of a larger number of genes/proteins. Likewise, in drug development, high throughput allows for the screening of large chemical libraries for desired effectiveness. In cytometry, increased throughput allows larger cell populations to be screened and sorted more quickly.
However, a need remains for a high throughput hyperspectral imaging flow cytometer that enables large complex cell populations to be screened and sorted quickly.